A docking model based on mass spectrometric and biochemical data describes phage packaging motor incorporation

Chi Yu Fu, Charlotte Uetrecht, Sebyung Kang, Marc Morais, Albert J R Heck, Mark R. Walter, Peter E. Prevelige

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

The molecular mechanism of scaffolding protein-mediated incorporation of one and only one DNA packaging motor/connector dodecamer at a unique vertex during lambdoid phage assembly has remained elusive because of the lack of structural information on how the connector and scaffolding proteins interact. We assembled and characterized a φ29 connector-scaffolding complex, which can be incorporated into procapsids during in vitro assembly. Native mass spectrometry revealed that the connector binds at most 12 scaffolding molecules, likely organized as six dimers. A data-driven docking model, using input from chemical cross-linking and mutagenesis data, suggested an interaction between the scaffolding protein and the exterior of the wide domain of the connector dodecamer. The connector binding region of the scaffolding protein lies upstream of the capsid binding region located at the C terminus. This arrangement allows the C terminus of scaffolding protein within the complex to both recruit capsid subunits and mediate the incorporation of the single connector vertex.

Original languageEnglish (US)
Pages (from-to)1764-1773
Number of pages10
JournalMolecular and Cellular Proteomics
Volume9
Issue number8
DOIs
StatePublished - 2010

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Bacteriophages
Product Packaging
Packaging
Capsid
Proteins
DNA Packaging
Mutagenesis
Dimers
Mass spectrometry
Mass Spectrometry
Molecules
DNA

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Analytical Chemistry
  • Medicine(all)

Cite this

A docking model based on mass spectrometric and biochemical data describes phage packaging motor incorporation. / Fu, Chi Yu; Uetrecht, Charlotte; Kang, Sebyung; Morais, Marc; Heck, Albert J R; Walter, Mark R.; Prevelige, Peter E.

In: Molecular and Cellular Proteomics, Vol. 9, No. 8, 2010, p. 1764-1773.

Research output: Contribution to journalArticle

Fu, Chi Yu ; Uetrecht, Charlotte ; Kang, Sebyung ; Morais, Marc ; Heck, Albert J R ; Walter, Mark R. ; Prevelige, Peter E. / A docking model based on mass spectrometric and biochemical data describes phage packaging motor incorporation. In: Molecular and Cellular Proteomics. 2010 ; Vol. 9, No. 8. pp. 1764-1773.
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